Anti-tumor phosphatase and tensin homolog deleted on chromosome ten (PTEN)-VP22 gene

A PTEN-VP22, anti-tumor technology, applied in the field of genetic engineering, can solve the problems of low PTEN gene transduction efficiency, hindering the clinical application of PTEN gene therapy, etc.

Inactive Publication Date: 2014-12-03
川北医学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing methods of directly applying the PTEN gene to vectors or plasmids and performing gene therapy on tumor cells, due to the low transduction efficiency of the PTEN gene, have become one of the reasons that hinder the clinical application of PTEN gene therapy for tumors

Method used

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  • Anti-tumor phosphatase and tensin homolog deleted on chromosome ten (PTEN)-VP22 gene
  • Anti-tumor phosphatase and tensin homolog deleted on chromosome ten (PTEN)-VP22 gene
  • Anti-tumor phosphatase and tensin homolog deleted on chromosome ten (PTEN)-VP22 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1—construct anti-tumor PTEN-VP22 gene by way of PCR

Embodiment 2

[0072] The PTEN-VP22 gene of the control example 1-3 obtained in embodiment 2-embodiment 1 is for the cell in vitro experiment of tumor cell

[0073] The PTEN-VP22 gene of Control Example 1-3, as well as PTEN and VP22 genes were connected into eukaryotic expression plasmid pcDNA3 to construct pcDNA3-PTEN-VP22, pcDNA3-PTEN and pcDNA3-VP22 expression vectors respectively.

[0074] Cell experiment 1: Inhibitory effect on breast cancer cell line BT549

[0075] A. Time-effect relationship: Transfect 1 μg of pcDNA3, pcDNA3-PTEN, pcDNA3-VP22, and pcDNA3-PTEN-VP22 plasmids into BT549 cells cultured in a 24-well plate, and set up a negative control group of untransfected BT549 cells. 8 replicate wells. Cells were cultured for 4 h after transfection, and 100 μl of cell suspension (1×10 3 cells), at 37°C, 5% CO 2 Cultivate under conditions; after 24h, 48h, 72h, 84h, and 96h, add 10 μl of CCK-8 solution to each well at 37°C, 5% CO 2 After incubation for 2 hours, the A450 value was mea...

Embodiment 3

[0118] The PTEN-VP22 gene of control example 1 obtained in embodiment 3-embodiment 1 is to the animal experiment of tumor cell

[0119] The PTEN-VP22 gene and PTEN of Control Example 1 were connected into the eukaryotic expression plasmid pcDNA3 to construct pcDNA3-PTEN-VP22 and pcDNA3-PTEN expression vectors respectively.

[0120] Animal Experiment 1: Inhibitory Effect on Breast Cancer Cells

[0121] Breast cancer cells BT549 were inoculated subcutaneously in nude mice to establish tumor-bearing nude mice, and the tumor-bearing nude mice were randomly divided into three groups: experimental group 1, experimental group 2 and negative control group. Experimental group 1 was injected intratumorally with 500uL, 100μg / mL pcDNA3-PTEN-VP22 gene expression vector, experimental group 2 was injected with 500uL, 100μg / mL pcDNA3-PTEN gene expression vector, and the negative control group was not injected. Observe the survival rate and average tumor volume of the three groups of mice on ...

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Abstract

The invention relates to the field of gene engineering and concretely relates to an anti-tumor phosphatase and tensin homolog deleted on chromosome ten (PTEN)-VP22 gene. The anti-tumor PTEN-VP22 gene comprises a nucleotide sequence (i) shown in the formula SEQ ID NO.1 in the sequence table or comprises a nucleotide sequence (ii) having homology of 95% with the nucleotide sequence (i). A protein coded by the anti-tumor PTEN-VP22 gene can effectively inhibit tumor cells, can effectively penetrate a cell membrane thereby producing inhibition effects on tumor cells and has active effects in PTEN gene clinical application.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to an anti-tumor PTEN-VP22 gene. Background technique [0002] Tumor is an important disease that threatens human health, and the research on its treatment has always been the focus of medical and life science research. Tumor biotherapy has been recognized by the world medical community as the fourth major treatment method after surgery, radiotherapy and chemotherapy, and gene therapy is one of the most active research fields in tumor biotherapy. The principle of gene therapy is to introduce the target gene into target cells using gene transfer technology, so that the target cells can express the gene to obtain specific functions, and then execute or mediate the killing and inhibition of tumors, so as to achieve the purpose of treatment. [0003] Oncogene activation and tumor suppressor gene inactivation are currently recognized as important molecular mechanisms of tumorigenesis....

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/63C12N15/10A61K48/00A61P35/00C07K14/47
Inventor 雷军余娴胥正敏李婷婷
Owner 川北医学院
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